Liu Yong-Qi, Li Yi, Qin Jie, Wang Qian, She Ya-Li, Luo Ya-Li, He Jian-Xin, Li Jing-Ya, Xie Xiao-Dong
Key Laboratory of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou University, Lanzhou, China E-mail :
Asian Pac J Cancer Prev. 2014;15(5):2169-77. doi: 10.7314/apjcp.2014.15.5.2169.
Matrine, a main active component extracted from dry roots of Sophora flavecens , has been reported to exert antitumor effects on A549 human non-small lung cancer cells, but its mechanisms of action remain unclear. To determine effects of matrine on proliferation of A549 cells and assess possible mechanisms, MTT assays were employed to detect cytotoxicity, along with o flow cytometric analysis of DNA content of nuclei of cells following staining with propidium iodide to analyze cell cycle distribution. Western blotting was performed to determined expression levels of Bax, Bcl-2, VEGF and HDAC1, while a microarray was used to assessed changes of miRNA profiles. In the MTT assay, matrine suppressed growth of human lung cancer cell A549 in a dose- and time- dependent manner at doses of 0.25-2.5 mg/ml for 24h, 48h or 72h. Matrine induced cell cycle arrest in G0/G1 phase and decreased the G2/M phase, while down-regulating the expression of Bcl2 protein, leading to a reduction in the Bcl-2/Bax ratio. In addition, matrine down regulated the expression level of VEGF and HDAC1 of A549 cells. Microarray analysis demonstrated that matrine altered the expression level of miRNAs compared with untreated control A549 cells. In conclusion, matrine could inhibit proliferation of A549 cells, providing useful information for understanding anticancer mechanisms.
苦参碱是从苦参干燥根中提取的主要活性成分,据报道其对人非小细胞肺癌A549细胞具有抗肿瘤作用,但其作用机制尚不清楚。为了确定苦参碱对A549细胞增殖的影响并评估可能的机制,采用MTT法检测细胞毒性,并用碘化丙啶染色后通过流式细胞术分析细胞核DNA含量以分析细胞周期分布。进行蛋白质免疫印迹法以测定Bax、Bcl-2、VEGF和HDAC1的表达水平,同时使用微阵列评估miRNA谱的变化。在MTT试验中,苦参碱在0.25-2.5mg/ml剂量下作用24小时、48小时或72小时,以剂量和时间依赖性方式抑制人肺癌细胞A549的生长。苦参碱诱导细胞周期停滞在G0/G1期并减少G2/M期,同时下调Bcl2蛋白的表达,导致Bcl-2/Bax比值降低。此外,苦参碱下调A549细胞中VEGF和HDAC1的表达水平。微阵列分析表明,与未处理的对照A549细胞相比,苦参碱改变了miRNA的表达水平。总之,苦参碱可以抑制A549细胞的增殖,为理解其抗癌机制提供有用信息。